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. 2001 Apr 17;98(10):5815–5820. doi: 10.1073/pnas.081612998

Figure 2.

Figure 2

Relation between cholesterol, membrane fluidity, and α-secretase activity. (A) Secretion of APPsα as a function of cellular cholesterol amount and of fluorescence anisotropy in membranes of HEK cells. HEK cells were incubated with 10 mM MβCD for various times (0, 2, 5, 10, 20, 30 min) to remove defined cholesterol amounts. After 4 h, the medium was collected, and secreted APPsα was identified as described in Fig. 1. The cell pellet was split for determination of cellular cholesterol and protein content and for measurements of 1,6-diphenyl-1,3,5-hexatriene steady-state fluorescence anisotropy in cell membranes. The data are expressed as percentage of control and are the averages ± SD of at least three experiments. (B) Influence of steroids on secretion of APPsα from HEK ADAM 10 cells, which were incubated in the presence of 10 mM MβCD for 30 min at 37°C. Subsequently, cells were incubated in the presence of 0.3 mM steroid–MβCD complexes for 30 min at 37°C.